Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein

doi:10.1186/s12934-019-1100-6

CORC > 海洋研究所 > 中国科学院海洋研究所 > 实验海洋生物学重点实验室

	Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein
	Chen, Huaxin 1,2,3; Jiang, Peng 1,2,3
刊名	MICROBIAL CELL FACTORIES
	2019-03-20
卷号	18 页码:13
关键词	Phycobiliprotein Phycobilin Chromophorylation ratio Plasmid stability E. coli
ISSN号	1475-2859
DOI	10.1186/s12934-019-1100-6
通讯作者	Chen, Huaxin(chhx@qdio.ac.cn)
英文摘要	BackgroundPhycobiliproteins (PBPs) are light-harvesting protein found in cyanobacteria, red algae and the cryptomonads. They have been widely used as fluorescent labels in cytometry and immunofluorescence analysis. A number of PBPs has been produced in metabolically engineered Escherichia coli. However, the recombinant PBPs are incompletely chromophorylated, and the underlying mechanisms are not clear.Results and discussionIn this work, a pathway for SLA-PEB [a fusion protein of streptavidin and allophycocyanin that covalently binds phycoerythrobilin (PEB)] biosynthesis in E. coli was constructed using a single-expression plasmid strategy. Compared with a previous E. coli strain transformed with dual plasmids, the E. coli strain transformed with a single plasmid showed increased plasmid stability and produced SLA-PEB with a higher chromophorylation ratio. To achieve full chromophorylation of SLA-PEB, directed evolution was employed to improve the catalytic performance of lyase CpcS. In addition, the catalytic abilities of heme oxygenases from different cyanobacteria were investigated based on biliverdin IX and PEB accumulation. Upregulation of the heme biosynthetic pathway genes was also carried out to increase heme availability and PEB biosynthesis in E. coli. Fed-batch fermentation was conducted for the strain V5ALD, which produced recombinant SLA-PEB with a chromophorylation ratio of 96.7%.ConclusionIn addition to reporting the highest chromophorylation ratio of recombinant PBPs to date, this work demonstrated strategies for improving the chromophorylation of recombinant protein, especially biliprotein with heme, or its derivatives as a prosthetic group.
资助项目	development funds of science and technology of Shinan district, Qingdao[2016-3-010-ZH] ; National Natural Science Foundation of China[41276164]
WOS研究方向	Biotechnology & Applied Microbiology
语种	英语
出版者	BMC
WOS记录号	WOS:000462299800001
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/154968]
专题	海洋研究所_实验海洋生物学重点实验室
通讯作者	Chen, Huaxin
作者单位	1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266071, Peoples R China 3.Chinese Acad Sci, Ctr Ocean Mega Sci, Qingdao 266071, Peoples R China
推荐引用方式 GB/T 7714	Chen, Huaxin,Jiang, Peng. Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein[J]. MICROBIAL CELL FACTORIES,2019,18:13.
APA	Chen, Huaxin,&Jiang, Peng.(2019).Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein.MICROBIAL CELL FACTORIES,18,13.
MLA	Chen, Huaxin,et al."Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein".MICROBIAL CELL FACTORIES 18(2019):13.